A Novel Heterozygous Intronic Mutation in the FBN1 Gene Contributes to FBN1 RNA Missplicing Events in the Marfan Syndrome

Figure 3

Western blot analysis of minigene-derived proteins. (a) FLAG/Myc-tagged reference (FBN1-Ref) and mutant (FBN1-Mut) minigene plasmids (PL) were transfected into HeLa cells as indicated at the top of each lane, and cell lysates were analyzed by Western blotting with a mouse monoclonal anti-FLAG (upper panel) or anti-Myc (lower panel) antibody. The results from experiments performed on four batches of cells for each transfection are shown. Lane 13: cells transfected with empty vector. The Western blot detection of the FLAG-tagged bacterial alkaline phosphatase (BAP) was used as a marker of equivalent transfection efficiency and equal loading. MW values (kDa) of the bands detected are shown in brackets. (b) COS-7 cells were transfected as indicated at the top of each lane and analyzed by Western blotting with a mouse monoclonal anti-FLAG antibody (upper panel). Protein load (lower panel): membrane stained with Amido Black 10B after immunostaining. Other indications as in (a).

(a)

(b)